Fig. 1. Ercc1 expression in mouse testis. (A) Pattern of Ercc1 expression in mouse tissues. Upper panel: total RNA (30 µg) from a range of tissues was analysed by northern blotting using an Ercc1 cDNA probe [Probe a in Selfridge et al. (Selfridge et al., 2001)]. Lower panel: total protein (80 µg) was analysed by western blotting using an antibody raised against a fragment of mouse Ercc1. (B) Developmental pattern of Ercc1 mRNA in testis. Total RNA (30 µg) extracted from testes of mice (ranging from 8-42 days post partum) was analysed by northern blotting using an Ercc1 cDNA probe. The filter was then reprobed for actin mRNA using a mouse -actin cDNA probe (Minty et al., 1981). The Ercc1/actin mRNA ratio was determined by phosphorimagery and is expressed relative to the p8 sample. (C) Developmental pattern of Ercc1 protein in testis. Total protein (80 µg) extracted from testes of mice (ranging from 8-35 days post partum) was analysed by western blotting using the antibody against Ercc1. Wild type (Wt) and Ercc1 null liver samples were used to demonstrate the specificity of the antibody. The filter was then reprobed with an antibody against mitochondrial core protein II (CP II). The Ercc1/CP II protein ratio was determined by densitometry and is expressed relative to the p8 sample. The ratios shown are the means of two separate determinations on two independent samples at each age. (D) Analysis of Ercc1 transgene expression. Upper panel: total RNA (30 µg) extracted from a range of tissues from transgene-positive Ercc1-deficient mice was analysed by northern blotting using a probe from the 5' end of the mouse transthyretin (Ttr) gene [Probe b in Selfridge et al. (Selfridge et al., 2001)]. mRNA from the endogenous Ttr gene and from the transgene (TG) is indicated. Lower panel: total protein (80 µg) was analysed by western blotting using the antibody against Ercc1.

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